Research ArticleEthnoecological, Elemental, andPhytochemical Evaluation of FivePlant Species of Lamiaceae in Peshawar, Pakistan

Sulaiman ,1 S. M. Shah,2 Sadaf,1 M. Amin,1 B. Gul,1 and M. Begum1

1Department of Botany, University of Peshawar (UoP), Peshawar, KP 25120, Pakistan2Centre of Plant Biodiversity, University of Peshawar (UoP), Peshawar, KP 25120, Pakistan

Correspondence should be addressed to Sulaiman; sulaiman097@uop.edu.pk

Received 28 May 2020; Revised 16 July 2020; Accepted 23 July 2020; Published 27 August 2020

Academic Editor: Umit Tursen

Copyright © 2020 Sulaiman et al. +is is an open access article distributed under the Creative Commons AttributionLicense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work isproperly cited.

+e use of medicinal plants as an unconventional health treatment is gaining considerable recognition and popularity worldwide.+e current study was designed to inspect five medicinally important species (such as Mentha longifolia (L.) Huds., Menthapiperita L., Mentha spicata L., Ocimum basilicum L., and Rosmarinus officinalis L.) of Lamiaceae, collected from district Peshawar,through ethnoecological, phytochemical, and elemental analyses. Biological spectra expressed that therophytes (60%) were thedominant life-form class, while nanophyll (60%) was the leading class among leaf size. +e ethnobotanical profile showed that allthe species were medicinal and ornamental (100%) each, while 60% were used in spices. Quantitative analysis for the macro- andmicrominerals confirmed the presence of 13 elements (C, N, O, Mg, K, P, S, Ca, Al, Si, Fe, Cl, and Na), which were present invarying amounts from species to species. +e methanol extract of leaf samples was used for the analysis of phytochemicalconstituents such as saponins, flavonoids, tannins, terpenoids, phlobatannins, steroids, and anthraquinones. +e medicinalpotential of these plants was correlated with the presence of these phytochemicals. Due to the presence of active constituents, theplants had high potential in antifungal, antidiuretic, antioxidant, and anti-inflammatory activities.

1. Introduction

Peshawar is the capital city of Khyber Pakhtunkhwa,province of Pakistan, located at the altitude of 300 metersbetween 35°50′37N latitude and 71°21′45E longitude (Fig-ure 1). Its population is 1,970,042, and it covered an area of1,257 kilometer square [1]. Edaphology deals with the in-fluence of soil on living things specially plants. Peshawar iscovered with the combined deposits of sand, silt, and loamysoil, which support a great diversity of flora. Peshawar is notlocated in the monsoon region unlike the other northernparts of Pakistan. +e area has extreme conditions; thecoldest month is January with average maximum temper-ature 18.35°C, and the hottest month is June having a meanmaximum temperature of 40.8°C. +e relative humidityvaries from 46% in June to 76% in August [2]. +e averageannual rainfall recorded was 454.2mm. Plants are used as

medicine and food since time immemorial round the globedue to their most valuable properties. Medicinal plants playan important role in drug discovery, and human beings usedthem for various purposes from ancient time.

About 80% of the population in the developing countriesdepends onmedicinal plants for primary health care [3]. Outof 50,000 angiospermic plants used as medicines, more than600 species are used in Pakistan for curing various diseases[4, 5]. +e family Lamiaceae, also known as Labiatae or mintfamily, consists of highly valued medicinal plants withcosmopolitan distribution and worldwide source of spicesand various extracts [6]. Within this family, about 236genera and more than 6000 species are present in whichgenera Ocimum and Mentha provide various taxa [6, 7].Ethnoecological studies have been carried out by variousresearch studies in Peshawar and adjoining areas [8–14].+eplants of this family contain active chemical constituents and

HindawiScientificaVolume 2020, Article ID 2982934, 8 pageshttps://doi.org/10.1155/2020/2982934

secondary metabolites like vitamins and minerals [15].Rosmarinus officinalis is used as an antimicrobial, anti-inflammatory, antidiuretic, antidiabetic, and anticanceragent [16]. Mentha species possess antioxidant propertiesbecause of the presence of active components, such asrosmarinic acid, menthol, carvone, menthone, and fla-vonoids [17]. Minerals play a primary role in reproduc-tion, growth, health, and proper functioning of livingorganisms [18]. Elemental analysis showed that membersof Lamiaceae taxa are rich in macro- and microelements.Phytochemicals are naturally occurring biologically activecompounds found in plants, which are liable for healthbenefits of humans [19, 20]. +e phytochemical screeningof various plants of Lamiaceae was carried out by Asghariet al., Cocan et al., and Mahendra and Kakde [21–23] andelemental analysis by Arika et al. and Gogoasa et al.[24, 25].

+e aim of the present work was to evaluate the macro-and microelements quantitatively and phytochemical con-stituents qualitatively. +e ecological, ethnobotanical, andethnomedicinal properties were also known. +e findingswill help in future research studies.

2. Materials and Methods

2.1.CollectionofPlants. Fresh leaves of five medicinal plants,M. longifolia, M. piperita, M. spicata, O. basilicum, andR. officinalis, were collected from district Peshawar. Plantspecimens were taxonomically identified in the Centre ofPlant Biodiversity by Dr. Syed Mukaram Shah (PhD), andvoucher specimens were deposited in the Herbarium ofDepartment of Botany, University of Peshawar.

2.2. Ecological and Ethnobotanical Profiles. Ecologicalcharacteristics of plants depend upon altitude, climate, andrelated environmental conditions. Life form, leaf size, leafshape, and phenology of the plants were observed [26–28].Ethnobotany is the cultural relationship among plants,people, and environment. Ethnobotanical information wasgathered through the literature and from the local inhabi-tants. Ethnomedicinal information was documented fromlocal hakims and aged people.

2.3. Phytochemical Analysis. Leaves were shade-dried forthree weeks, grinded into powder using an electrical grinder,and kept in polythene bags with proper labeling for furtheruse. 50 grams of each powder sample were added separatelyinto 250ml of methanol solvent in a conical flask at roomtemperature. After 48 hours, the extracts were filtered withWhatman No. 1 filter paper. +e extracts were stored in air-tight glass at 4°C for further analysis.

In test tubes, 0.5 gram of each plant sample was shakenwith 5ml of distilled water. Frothing which persists onwarming was taken as preliminary evidence for the presenceof saponins [29].

For flavonoid detection, 5ml of solution of dilute am-monia was mixed with 0.5 gram of the plant sample followedby the addition of concentrated sulphuric acid. Yellowcoloration indicates the presence of flavonoids which dis-appear later on standing [30].

For the detection of terpenoids, 3ml of concentratedsulphuric acid and 2ml of chloroform were added to 0.5gram of the plant sample to form a layer. Reddish-browncoloration showed the presence of terpenoids [31].

Peshawar city

Peshawar

Figure 1: Map of the study area. Source: Geography Department, University of Peshawar, KP, Pakistan.

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For steroid detection, 10ml of chloroform was mixed with1ml of the plant extract, and equal volume of concentratedH2SO4 was added by the side of the test tube. +e upper filmturns red, and the H2SO4 layer shows yellow with green flo-rescence. +is indicates the presence of steroids [32].

0.2 gram of each plant powder was boiled in one percentaqueous HCl solution. +e formation of red precipitateindicates the presence of phlobatannins [30, 31].

For the detection of tannins, 0.5 gram of each sample wasstirred with 100ml of distilled water, filtered, and 0.1 percent of

Elemental assessment

SEM coupled with EDXStage

Stub

Powder

Leaves

Elemental assessment

SEM coupled with EDX

Stub

Leaves

Display

Sample holder

Figure 2: Elemental analysis.

Table 1: Biological spectra of plants of Lamiaceae in Peshawar, Pakistan.

S. no. Plant species Voucher no. Life form Leaf size Leaf shape Phenology(A) Angiosperm(a) Dicots(1) Family Lamiaceae(1) Mentha longifolia (L.) Huds. B.Sul.015.UOP G Mic S S1(2) Mentha piperita L. B.Sul.016.UOP + N S S1(3) Mentha spicata L. B.Sul.017.UOP + N S S1(4) Ocimum basilicum L. B.Sul.018.UOP + Mic S S2(5) Rosmarinus officinalis L. B.Sul.019.UOP Ch N S S3Ch� chamaephytes, +� therophytes, G� geophytes, Mic�microphyll, N�nanophyll, Mes�mesophyll, S� simple, S1� pre-reproductive,S2� reproductive, and S3� postreproductive.

60%20%

20%

Therophytes

Geophytes

Chamaephytes

Figure 3: Life-form classes.

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ferric chloride reagent was added to the 20ml filtrate. +eappearance of blue-green coloration shows the presence oftannins [33].

For anthraquinone detection, 0.5 gram of the plantextract was shaken with 5ml of chloroform. +e solutionwas filtered, and 10 percent ammonia solution was addedto the filtrate. +e mixture was shaken thoroughly, andthe formation of pink/violet color in the ammonicalphase indicates the presence of anthraquinones [31].

2.4. Elemental Analysis. Plants were collected, shade-dried,and ground to powder. For the quantitative analysis ofmacro- and microelements, energy-dispersive X-ray spec-trometer (Model Perkin Elmer AA Analyst 700) was usedwhich is installed in the Centralized Resource Laboratory,University of Peshawar. 0.5 gram of each plant powder wasplaced on stub, which was grip in the sample holder. +esample holder was laid inside the stage, and the elements’peaks were observed through display (Figure 2).

Table 2: Summary of biological spectra of plant species of Lamiaceae in Peshawar, Pakistan.

S. no. Life-form classes No. of species Percentage (%)(A) Life form(i) +erophytes (+) 3 60(ii) Chamaephytes (Ch) 1 20(iii) Geophyte (G) 1 20Total 5 100(B) Leaf sizes(i) Nanophyll 3 60(ii) Microphyll 2 40Total 5 100(C) Leaf shape(i) Simple 5 100Total 5 100(D) Phenology(i) Pre-reproductive (S1) 3 60(ii) Reproductive (S2) 1 20(iii) Postreproductive (S3) 1 20Total 5 100

Table 3: Ethnobotanical profile of plant species of Lamiaceae in Peshawar, Pakistan.

S. no. Plant species (1) (2) (3)(1) Mentha longifolia (L.) Huds. + − +(2) Mentha piperita L. + − +(3) Mentha spicata L. + + +(4) Ocimum basilicum L. + + −

(5) Rosmarinus officinalis L. + + −

Total 5 3 3Percentage (%) 100 60 60(1)�medicinal, (2)� ornamental, and (3)� spices.

Table 4: Ethnomedicinal uses of plants of Lamiaceae in Peshawar, Pakistan.

S. no. Plant speciesPart used

Constituents DiseasesRoot Stem Leaves Flower

(1) Mentha longifolia(L.) Huds. + + + + Pulegone, menthone, borneol, and

flavonoids.Carminative stimulant, antispasmodic,

and headache.

(2) Mentha piperita L. − − + + Menthol, menthone, methylacetate, and steroids.

Irritable bowel syndrome, muscle pain,nerve pain, and itching.

(3) Mentha spicata L. − − + −Terpenoid, carvone, menthol,

menthone, and tannins.Antitumor, antioxidant, and

antimicrobial.

(4) Ocimum basilicumL. + − + + Saponins, coumarins,

anthocyanin, and terpenoids. Antioxidant, antifungal, and antiviral.

(5) Rosmarinusofficinalis L. − − + −

Rosmarinic acid, camphor,flavonoids, and saponins.

Antioxidant, antidiuretic, and anti-inflammatory.

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Quantitative results obtained were copied to an Excelworksheet.+e worksheet was referred to as raw data. All thedata in the raw data worksheet were copied to an editedworksheet where unnecessary data columns were deleted.Also, unnecessary element row lines were removed, leavingthe calculated averages as the final concentration data.

3. Results and Discussion

3.1. Ecological and Ethnobotanical Profiles. During thepresent research work, 5 plant species of Lamiaceae werecollected from the study area which were investigated fortheir various biological aspects such as phenology, life form,leaf shape, leaf size, and ethnobotanical uses along withphytochemical and elemental screening. During collectiontime, the phenological stage of Mentha species was pre-reproductive, while Ocimum and Rosmarinus were at re-productive and postreproductive stages (Table 1). Sixtypercent species were therophytes, and 20% were eachchamaephytes and geophytes (Figure 3). +e biological leaf

spectra showed that 60% species had nanophyllous and 40%had microphyllous leaves (Table 2). All the species had asimple leaf shape. Ethnobotanical profile showed that amajority of plant species were used in spices and had me-dicinal and ornamental importance (Table 3). Various partsof the plants contained active constituents such as menthol,menthone, and rosmarinic acids which are used as anti-fungal, antioxidant, anti-inflammatory, and antidiureticagents (Table 4). +e present work agrees with many otherresearch studies [17, 34], where both reported that thesespecies were used as antioxidant, antidiuretic, and antimi-crobial agents.

3.2. Macro- and Microelements’ Assessment. All the plantspecies had high mineral contents and can be used as a goodsource for medicines (Table 5). A total of 13 different ele-ments were quantitatively analyzed in all the five plantspecies. Among the major elements, carbon was maximum(70.25%) in R. officinalis and minimum (56.2%) in

Table 5: Mineral contents in plant species of Lamiaceae in Peshawar, Pakistan.

S. no. Plant speciesMinerals (%)

Major elements Minor elementsC N O Mg K P S Ca Al Si Fe Cl Na

(1) Mentha longifolia (L.) Huds. 60.19 7.74 34.07 0.37 5.90 0.40 0.61 2.30 — 0.61 — 0.54 —(2) Mentha piperita L. 56.26 7.89 36.42 0.38 4.01 0.59 0.40 1.53 0.17 0.54 0.24 0.50 —(3) Mentha spicata L. 56.69 8.23 33.90 0.39 3.41 0.59 0.76 2.01 0.31 0.93 0.29 1.09 0.39(4) Ocimum basilicum L. 57.59 9.78 33.72 0.58 4.46 0.46 0.52 2.67 0.25 0.75 0.17 0.89 —(5) Rosmarinus officinalis L. 70.25 — 29.91 0.27 2.15 0.27 0.28 1.05 0.14 0.40 — 0.19 —

0

10

20

30

40

50

60

70

80

Mentha longifolia (L.)Huds.

Mentha piperitaL.

Mentha spicataL.

Ocimum basilicumL.

Rosmarinus officinalisL.

CNO

KPS

Mg

CaAlSi

FeClNa

Figure 4: Elemental assessment of plant species: (a) Mentha spicata L.; (b) Mentha longifolia (L.) Huds.; (c) Rosmarinus officinalis L.; (d)Mentha piperita L.; (e) Ocimum basilicum L.

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M. piperita, while only nitrogen was absent in R. officinalis.In minor elements, chlorine was found highest (1.09%) inMentha spicata and lowest (0.19%) in R. officinalis (Fig-ures 4 and 5). O. basilicum and M. longifolia had highcalcium content which plays a key role in bone formation[35].M. longifolia had the highest potassium concentration

which plays a key part of many enzymes’ synthesis andplays a vital role in the activation of growth of enzymes[36, 37]. Arika et al. [24] and Gogoasa et al. [25] had re-ported C, N, O, Mg, K, P, S, Ca, Al, Si, Fe, Cl, and Naelements in Mentha species, Ocimum species, and inRosmarinus species.

(a) (b)

(c) (d)

(e)

Figure 5: Mentha spp.

Table 6: Phytochemical screening of plants of Lamiaceae in Peshawar, Pakistan.

S. no. Plants species Flavonoids Steroids Tannins Terpenoids Saponins Phlobatannins Anthraquinones(1) Mentha longifolia (L.) Huds. + − − + + − −

(2) Mentha piperita L. + + + − + − −

(3) Mentha spicata L. − + + + − + −

(4) Ocimum basilicum L. + + − + + − +(5) Rosmarinus officinalis L. + − + + + − −

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3.3. Phytochemical Screening. +e phytochemical tests ofplant extracts were determined for the presence of flavo-noids, saponins, phlobatannins, tannins, steroids, terpe-noids, and anthraquinones (Table 6).+e results showed thatthe majority of the plants contained flavonoids, terpenoids,and saponins, while anthraquinones were only present inO. basilicum. +e extract of M. piperita and R. officinaliscontained flavonoids which are used in various ailmentsincluding dyspepsia, renal pain, arthritis, and antitumor[38]. Steroids were present in most plant extracts which areof great importance in pharmacy because they possesscompounds like sex hormones [39]. +e finding agrees withthat of Adham [6] and Inas et al. [32], who also observedflavonoids, phlobatannins, saponins, terpenoids, tannins,steroids, and anthraquinones in these species.

4. Conclusions

+e use of medicinal plants is a traditional practice inPakistan; thus, it is very important to evaluate the thera-peutic use of plants through scientific methods and provideinformation about the species that could be used in thefuture for their properties. +e medicinal plants appear to berich in secondary metabolites and mineral contents, widelyused in traditional medicine to combat and cure variousdiseases. +e selected five species in this study consist ofmany useful phytochemical compounds and active elementshaving important biological properties. Phytochemical in-vestigation and elemental analysis of M. longifolia,M. piperita, M. spicata, O. basilicum, and R. officinalisshowed significant results and may be used for curing ofdifferent ailments and manufacturing of new drugs in thefuture.

Data Availability

+e data used to support the findings of this study are in-cluded within the article.

Conflicts of Interest

+e authors declare that there are no conflicts of interestregarding the publication of this paper.

Acknowledgments

+e authors are indebted to Dr. Syed Mukaram Shah ofCentre of Plant Biodiversity, University of Peshawar,Peshawar, for his supervision and proofreading the article.

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